Part:BBa_K4645004

VqmAphage

vqmA gene would translate into protein VqmA_Phage.
Vibrio cholerae uses a quorum-sensing (QS) system composed of the autoinducer 3,5-dimethylpyrazin-2-ol (DPO) and receptor VqmA, which together repress genes for virulence and biofilm formation.[1] VqmA_Vc activates expression of the vqmR gene encoding the small regulatory RNA VqmR. Vibriophage VP882 encodes a VqmA homolog called VqmA_Phage that activates the qtip promoter (Pqtip). It’s worth noting that, VqmA_Phage can activate vqmR expression but VqmA_Vc cannot activate expression of qtip. Promoter selectivity is driven by each VqmA DNA-binding domain and key DNA sequences in the vqmR and qtip promoters are required to maintain specificity.[2]

Characterization

Successful protein expression

We transformed vqmA into BL21 and cultivated in 37℃, 220 rpm. When the OD600 of bacteria reached 0.6, we induced bacteria with 1mM, 2mM IPTG, then cultivated for about 12 hours. Here is SDS-PAGE gel figure of vqmA.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 535
  
• 1000
  COMPATIBLE WITH RFC[1000]

Reference

[1] Silpe JE, Bassler BL. A Host-Produced Quorum-Sensing Autoinducer Controls a Phage Lysis-Lysogeny Decision. Cell. 2019 Jan 10;176(1-2):268-280.e13.
[2] Duddy OP, Huang X, Silpe JE, Bassler BL. Mechanism underlying the DNA-binding preferences of the Vibrio cholerae and vibriophage VP882 VqmA quorum-sensing receptors. PLoS Genet. 2021 Jul 6;17(7):e1009550.